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WO2007081235A1 - Procede de traitement de maladies cancereuses - Google Patents

Procede de traitement de maladies cancereuses Download PDF

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Publication number
WO2007081235A1
WO2007081235A1 PCT/RU2006/000144 RU2006000144W WO2007081235A1 WO 2007081235 A1 WO2007081235 A1 WO 2007081235A1 RU 2006000144 W RU2006000144 W RU 2006000144W WO 2007081235 A1 WO2007081235 A1 WO 2007081235A1
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dna
cells
cell
gene
fragments
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Russian (ru)
Inventor
Mikhail Arkadievich Shurdov
Sergey Stanislavovich Bogachev
Leonid Anatolievich Yakubov
Vladimir Alexeevich Rogachev
Valery Petrovich Nikolin
Natalia Sergeevna Zhdanova
Nelly Alexandrovna Popova
Anastasia Sergeevna Likhacheva
Tamara Egorovna Sebeleva
Alexander Gennadievich Shilov
Ljudmila Vasilievna Mechetina
Oxana Vyacheslavovna Vratskikh
Sergey Nikolaevich Seregin
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Priority to JP2008551212A priority patent/JP2009523788A/ja
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/711—Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to medicine, in particular, to methods for treating patients with preparations containing genetic material, and can be used to treat patients with cancer, the cause of which are mutations in oncogenes, cancer suppressor genes and total homogenization of alleles of cell genes undergoing cancerous transformation.
  • the method is characterized by a relatively low treatment efficiency and limited use, since according to this method, mutations must be accurately determined before treatment begins.
  • SUBSTITUTE SHEET (RULE 26) The closest in essence to the proposed one is a method based on the activation of DNA repair in cells, in accordance with which delivery of repair enzymes to skin cells is performed in accordance with US patent No. 5,302,389, 04/12/1994.
  • the required task is to expand the scope and treatment of patients with cancer, the cause of which are both mutations in oncogenes and cancer suppressor genes, as well as total homogenization of genes.
  • the technical objective of the present invention is the development of a method for influencing cancer cells and the tumor as a whole in such a way that gene homeostasis changes when the cells undergo a reverse genetic degeneration in which the main disappears, experimentally confirmed on the culture of human adenocarcinoma cells and artificially induced mouse tumors
  • a property of a cancer cell is its unlimited proliferative activity.
  • the desired result is achieved by the fact that, according to the method of treating cancer based on the introduction of fragmented DNA into the body, fragments of homologous DNA are used that make up the whole genome of a physiologically and genetically healthy donor, while the amount of introduced DNA is equal to or greater than the amount of blood plasma DNA itself and tissue fluids of the patient, but not more than the maximum allowable amount of 30 m kg / ml.
  • SUBSTITUTE SHEET (RULE 26) genetically healthy DNA, the natural mechanism of its delivery and deposition in the interchromosomal space, and the homologous exchange of the delivered fragments and the corresponding sections of chromosomes carrying mutations that led to the oracle of this cell. Therefore, the proposal meets the criteria of novelty and inventive step.
  • the first sheet of the drawings shows the distribution of fragmented extracellular human DNA over the cell compartments of the cell culture of MCF-7 cells depending on the time it was in the culture medium (native conditions for phoresis, logarithmic phase of culture growth).
  • FIG. 1 and FIG. 2 cytoplasmic fraction, FIG. 3 and FIG. 4 - interchromosomal fraction, FIG. 5 and FIG. 6 - chromatin.
  • FIG. 1, FIG. 2, FIG. 3 agarose blocks stained with ethidium bromide.
  • FIG. 2, FIG. 4, FIG. 6 radiographs of the same blocks after drying.
  • the numbers above the blocks indicate the incubation time of the cell culture with labeled ⁇ 32 p extracellular DNA and ⁇ dATP *.
  • the numbers to the left and right of the blocks are molecular weight markers, etc. about. ⁇ is the original ⁇ dATP * precursor.
  • FIG. 7 shows the effect of the DNA preparation on the growth of Ehrlich’s cell tumor in terms of the change in the average weight of ascites in grams relative to
  • a healthy sequence and sequence of a specific region of the casnase 3 mRNA gene containing the repaired exon 4, exons 5 and 6 are also presented.
  • the figure shows: a set of primers and the expected PCR fragment of the caspase 3 gene of 431 bp. in the PT procedure; a set of primers and expected PCR fragments of 236 bp and 304 bp confirming the accession of fragment 431 bp .; amplification of the 431 bp fragment indicates
  • SUBSTITUTE SHEET (RULE 26) the existence of mRNA synthesized from the caspase 3 repaired gene; amplification of PCR fragments 236 bp and 304 bp, indicating the conformity of the internal structure of the fragment 431 bp the mRNA region of the caspase 3 gene containing the repaired exon 4, exons 5 and 6; comparison of the mRNA sequence of the caspase 3 gene containing exons 4, 5 and 6 and sequenced fragments 236 and 304 bp as internal segments obtained from a 431 bp fragment matrix
  • the proposed method for the treatment of cancer is implemented as follows.
  • fragments of homologous DNA are used that make up the complete genome of a physiologically and genetically healthy donor.
  • the amount of introduced fragmented DNA is equal to or exceeds the amount of the actual DNA of the blood plasma and tissue fluids of the patient, which is many times greater than the amount of DNA of blood plasma and tissue fluids of a healthy donor, but not more than the maximum allowable amount of 30 ⁇ g / ml.
  • DNA fragments having a biologically active size are delivered to actively dividing cells of the body, including cells that underwent cancer degeneration using the natural delivery mechanism (blood flow, specific receptors located on the surface of actively dividing cells having a high affinity for these DNA fragments).
  • DNA fragments delivered into the intracellular space are deposited in the interchromosomal compartment of the nucleus and are subjected to intranuclear processing.
  • a homologous, fragmented DNA when introduced into the body from a physiologically and genetically healthy donor, it is absorbed by actively dividing cancerous regenerated cells through the natural delivery mechanism inherent in actively dividing cells (receptor-mediated pinocytosis).
  • the fragmented DNA Delivered to the nuclear compartment — the interchromosomal space — the fragmented DNA is deposited and enters into the process of homologous exchange with the corresponding loci of the chromosomes.
  • DNA fragments deposited in the interchromosomal space replace mutant regions of genes, mutations in which led to cancerous cell degeneration into healthy ones, and also replace self-homogenized alleles, which is typical for oracular cells, leading them to a heterozygous state characteristic of a healthy cell.
  • cancer cells undergo partial or complete reversal degeneration and enter the path of development of a differentiated cell.
  • genomic DNA is a matrix of the body’s life that includes information about both the body as a whole and information about its development. Initially, genomic DNA is identical in all cells of the body. However, as the body grows, the genomic DNA of each cell undergoes mutational influences caused by environmental factors and errors that occur during cell replication and repair. It is known that
  • SUBSTITUTE SHEET (RULE 26) repair mechanisms replace incorrect or defective DNA bases and again close the ends of the DNA molecule after one- and two-link breaks (DCR) of chains immediately after mutational events. For this, they can use a second DNA strand as a template. In cases where the lesion sites are very long and affect both DNA strands, repair becomes problematic, and a mutation may result from the violation. Thus, mutagenic environmental factors and errors in cell repair and replication are sources of somatic mutations in cells. In the modern scientific understanding of the genesis of cancers, it is believed that the gradual accumulation of mutations in cells causes a multistage cancer transformation.
  • cancer treatment methods should be aimed at eliminating the cause of the disease, and, above all, at treating (correcting) mutations.
  • the only, fundamental approach to the treatment of diseases associated with genetic disorders, which is considered cancer, is a change in one way or another of the genome of the cells of the individual organism in the oracle and restoration of the original genetic homeostasis characteristic of a healthy cell.
  • numerous methods of gene therapy have been developed, however, all of them have serious shortcomings and are ineffective in the framework of a complex eukaryotic organism.
  • a gene-substitution strategy developed to overcome the problem of insertional mutagenesis on yeast and later on mammalian cells included the use of linear recombinant DNA in which the two ends are homologous to the regions flanking the replaced gene, and the gene itself is replaced by a selectable marker [Thoglas KR, Saresshi MR. Sit-direction mutagepsis bu gepe targetg ip mousem-derived couplings. CeII. 51 (3): 503-12. (1987)].
  • This type of gene targeting is called “ends-out” because the ends of the construct correspond to two diverging (from the target gene) chromosomal DNA sequences.
  • somatic GR in unicellular yeast is the main mechanism for precision repair of DCR, its effectiveness must be very high to ensure the salvation of the individual.
  • Early experiments with microinjections of plasmids into the cell nucleus [Saresshi, M. Gepertipg n ⁇ ice with target mutatiops. NATURE MEDICIPE 7, 1086-1090. (2001)] led to the discovery that plasmids were always integrated into the cellular genome in the form of a single concatemer consisting of all (often more than one hundred) injected plasmids assembled into a single molecule tail to head, and that the result was
  • SUBSTITUTE SHEET (RULE 26) surprisingly efficient operation of the GR mechanism.
  • the reason for the low efficiency of classical gene targeting involving the use of GH in mammalian cells may be the mismatch between the task of GH and its properties and the capabilities selected by evolution to perform its own, albeit unknown, function.
  • the data described in the literature allow us to identify some critical factors for the operation of the GR mechanism.
  • the efficiency of gene targeting obtained in these experiments may mean that GR prefers to work with either short fragments or forms short multimeric fragments that are most effective for use in recombination events using ligation of DNA fragments with ligase IV of the head-tail type.
  • a prerequisite for obtaining a recombinant product in mammalian cells is the length of the terminal homology of the fragments, which is about 200 bp (base pairs) [Lip Y., Lukasovix T., Weldmap A.S. Multirle ratthouses for DNA of DNA double-strapd breaks ip mamaliap chromosomes. MoI CeII Biol. 19, 8353-60 (1999); Nassif N., Engels W. DNA homologous protein for mit mitotis gift rerair ip Drorshila. Roc. Nail, Acad. Sretei. USA 90 (4): 1262-6 (1993)].
  • SUBSTITUTE SHEET (RULE 26) The ring forms of therapeutic plasmids practically do not participate in homologous exchange. Generation of DSB and formation of double-stranded ends of DNA molecule locally activates and amplifies chromosomal and extrachromosomal resombinatsiyu and gene targeting [Kusherlarati RS, Eves EM, Song KY, Morse BS, Smithies O. Nomologous resombipatiop betweep rlasmids Ip mammaliap sells glanders BE ephapsed bu treatmept ° F iprut DNA. Roc. Natl. Acad.Sci. USA 81, 3153-7 (1984)].
  • the authors by transfecting the cell with genetic constructs expressing synthetic zinc-finger nuclease, capable of specifically cleaving the sequence near the mutation in the IL2RD gene in the genomic DNA of cells and a plasmid containing a non-mutant fragment of the same gene that overlaps the SCID mutation region, showed a high, up to 20% level of gene correction.
  • Extracellular fragmented genomic DNA containing mutant sequences in proportions corresponding to the occurrence of specific mutations in the cells is formed as a result of programmed cell death or, in other words, apoptosis of body cells and is always present in the intercellular space and blood plasma [Apker P, Mulcahu H, Chen XQ , Stroun M. Detestsop of circulating tumor DNA and Tep Blood (Plasma / Srum) of sapseritis. Sapser Metastasis Rev. 18, 65-73 (1999); Giacona M. B. Ruben G. C, Iszkowski K. Roots T., Rogter D., Sorenson G.
  • the delivered fragments recombine with genomic DNA in the nuclei by the mechanism of homologous recombination (GR) and are capable of initiating this process themselves.
  • GR homologous recombination
  • allelic genes usually have a high degree of homology and often differ only in relatively small regions of altered sequences
  • another aspect of the proposed mechanism could be the replacement of allelic gene sequences in the chromosome with alternative ones, similar to replacing mutant sequences with non-mutant ones.
  • the complete genome of the individual is introduced into the body in the form of a set of polynucleotide molecules, which are fragments of genomic DNA, circulated and captured by cells, into the nucleus, where it is deposited and homologous recombination with the genomic DNA of the cell.
  • fragments of homologous DNA are used that make up the whole genome of a physiologically and genetically healthy individual in association with proteins of the nuclear matrix.
  • the size of exogenous fragments corresponds to the size of the fragments circulating in the blood plasma of the treated organism that arise during apoptotic nucleolysis (1-30 nucleosome units).
  • the prototypes use various artificial DNA constructs that do not make up the entire genome of the individual. This DNA is not in association with the proteins of the nuclear matrix and does not correspond to the size of the body's natural apoptotic DNA of 1-30 nucleosome units.
  • SUBSTITUTE SHEET (RULE 26) defined in tests for cytotoxicity performed on human cell culture (ip culture).
  • MCF-7 human mammary adenocarcinoma cell culture cells were cultured in RPMI 1640 medium with HMM L-glutamine and 50 ⁇ g / ml Sigma streptamycin (USA) at 37 ° C in the presence of 5% Biolot bovine serum (PBS) , ( Russia) in an atmosphere from 5% CO2 to a density of 0.7x107 cells per 4 wells in a 24 well plate.
  • PBS Biolot bovine serum
  • the number of MCF-7 human breast adenocarcinoma cell culture cells at the experimental point is 0.7 x 106. According to the protocol for CF-7, 107 cells contain 60 ⁇ g of DNA. One cell contains 6 picograms of DNA.
  • the total amount of DNA in the cells at the experimental point was 4.2 ⁇ g.
  • the amount of labeled DNA indicated in Table 1 was added to the experimental point.
  • the radioactivity count of the added DNA was the amount of cpm indicated in Table 1.
  • the radioactivity score of ⁇ dNTP * at the point was the amount of cpm indicated in Table 1.
  • the haploid genome of the human cell contains 3.3x10 9 bp.
  • the size of labeled genomic DNA fragments added to the medium was initially about 500 bp.
  • FIG. 1 shows an electrophoregram (right) and an X-ray film after exposure to the same agarose block after drying (left). Cytoplasmic fraction (upper group of blocks). DNA almost immediately penetrates the cytoplasm of cells. At the zero point, a low molecular weight fraction of mobility corresponding to the original labeled material is displayed. There is no label at the start of the zero point. In all other samples, a significant amount of labeled material is present at the start. This may mean that DNA forms high molecular complexes with the components of the cytoplasm. Obvious jellyfish precipitating DNA was not detected.
  • DNA almost immediately penetrates into the interchromosomal fraction of the nucleus.
  • a low molecular weight fraction of mobility corresponding to the original labeled material is displayed.
  • the entire high molecular weight fraction of DNA during centrifugation formed an obvious pellet that is not found in nuclear juice.
  • a change in the mobility of DNA is observed.
  • DNA forms a high molecular weight pool in the form of a “blurred cloud” rising from the lower part to the high molecular weight region of the agarose block.
  • a uniform cloud one can observe several discrete, size - dependent bands.
  • DNA banding can be seen in the chromatin fraction, where there is a slight contamination with interchromosomal material (the chromatin precipitate was not washed).
  • Such a pattern of the distribution of labeled material may indicate the following events. It is possible that fragmented DNA that has fallen into the nucleus acts as a primer primer for the synthesis of a DNA strand. This is expressed in the appearance of a different-sized set of labeled DNA fragments, which is detected on the phoresis as a “blurred - cloudy" labeled material reaching a size of about 10 etc. about. Another possibility is that DNA fragments are ligated at the ends. Such a conclusion can be drawn from the picture of discrete labeled bands, in size multiple of the original DNA. A combined variant of increasing the linear size of extracellular DNA in the interchromosomal space of the nucleus is also possible.
  • DNA (or a label in some other form) penetrates the nucleus almost immediately and integrates into the chromatin fraction of the nucleus. In the area of gel separation, a pronounced ladder of fragments in size multiple of the original DNA is observed. (The appearance of this labeled material is associated with a slight contamination of the interchromosomal fraction containing a high specific label).
  • ⁇ dATP * (in all blocks under the designation 180 ⁇ ). ⁇ dATP * is absent in the cytoplasmic fraction. However, there is a labeled fragment with a size that matches the size of the DNA that was used in the experiment.
  • the interchromosomal fraction In the interchromosomal fraction, it forms a similar, but less intense “blurred - cloud” morphologically completely corresponding to that for a similar fraction with labeled DNA. After 180 'the label is found in chromatin. Other points for ⁇ dATP * were not controlled. The amount of label that was taken in the culture medium at 180 minutes for labeled DNA and for ⁇ dATP * is approximately the same amount (see tables 1, 2). However, the amount of labeled material detected at the indicated experimental point with ⁇ dATP * is approximately b times less than in the case of DNA. Quantitative indicators of the behavior of labeled material.
  • ⁇ dATP * is delivered to all cell compartments not as a monomer but as a part of DNA fragments measuring about 500 bp in size. Further, this DNA is involved in all processes as well as DNA added to the medium. In all respects, the activity of ⁇ dATP * utilization is several times less intense than the utilization of labeled extracellular DNA. Further, it was found that extracellular DNA almost immediately (less than a minute if the cells are actively dividing) is delivered to all analyzed cell compartments (cytoplasm, nuclear space) and integrates into chromatin. Approximately the same amount of labeled DNA is present in the cytoplasm for all points except the zero point.
  • DNA is present in the interchromosomal space of the nucleus in the form of DNA fragments of various lengths.
  • the size of the fragments corresponds to the size of extracellular DNA added to the medium.
  • the linear size of DNA increases to sizes of the order of 10, i.e., about.
  • some fragments are ligated according to the "head to tail” type.
  • DNA processing denaturing conditions, data not shown
  • SUBSTITUTE SHEET (RULE 26) The possibility of correcting mutations by the action of extracellular DNA on cancer cells considered in the claimed method of treating cancers was investigated on the mutant caspase 3 gene of human adenocarcinoma. As a result of the studies, it was shown that the restoration of the activity of the caspase 3 gene of cells of human breast adenocarcinoma MCF-7, as well as the apoptotic pathway of development of previously cancer cells when exposed to extracellular DNA.
  • MCF-7 mammary adenocarcinoma cells and mouse sarcoma L929 were provided by the Vector Research Institute of Cell Cultures of the State Scientific Center of the World Bank "Vector”. Cells were cultured in RPMI 1640 medium with HMM L-glutamine and 50 ⁇ g / ml streptamycin (Sigma, USA) at 37 ° C in the presence of 5% fetal bovine serum (PBS) (Biolot, St. Russia) in an atmosphere with 5% CO2.
  • PBS fetal bovine serum
  • MCF-7 cells were cultured in RPMI 1640 medium with 10 mM L-glutamine and 50 ⁇ g / ml streptamycin in the presence of 5% fetal bovine serum (EBS) and 0.1 - 0.2 mg / ml fragmented human placenta DNA. Transfection time ranged from 5 to 40 days.
  • EBS fetal bovine serum
  • PCR analysis was carried out according to standard protocols using whales of the Medigen company ( Russia) and the BiRad company (USA). Real time PCR analysis was performed according to the protocol of the company "BioRad” (USA).
  • PCR DNA fragments were separated on a 2% agarose gel and stained with ethidium bromide. Isolation of RNA from cell culture.
  • RNA Purifieauitop Kit V-Gepe Biological Limit
  • SUBSTITUTE SHEET (RULE 26) The detection of either human caspase-3 enzyme or the corresponding mRNA with healed mutation in exon 4 in the cell extract of human DNA could be direct evidence of the homologous integration of the therapeutic fragment into the chromosome of mutant cells.
  • PT reverse transcriptase
  • RNA was isolated and cDNA synthesized from MCF 7 cell culture incubated with human DNA for 15 days.
  • primers 3 and 2 were used (Example 2, Pr. 3).
  • the desired fragment of 431 bp was obtained. including spliced exons 4 (without 20 initial nucleotides), exon 5 and exon 6.
  • the detection of this fragment in the total RNA fraction meant that the total RNA contains the mucosal gene of the capase 3 gene with a repaired 47 bp deletion.
  • Fragment 431 bp represents a cDNA copy of the mRNA region of the capase 3 gene containing spliced exons 4 (without 20 initial nucleotides, as determined by the primer site), exon 5 and exon 6.
  • the left edge of the fragment is 431 bp.
  • caspase gene is located on chromosome 4, has a size of 21751 bp and consists of 8 exons.
  • two products of the expression of a given gene can exist. These are transcripts of size 17 and 19, etc. about. and their protein products [.WoIf B.V., Schuller M., Echiverri F. and Green D. R .. Caspase-3 is the primary choice of the DNA complex of the DNA complex protein-45 / integrator compound. J. Biol. Chem. 274, 30651-30656 (1999)].
  • the appearance of the functional enzyme caspase 3 requires a complex process of reading mRNA in which 8 exons are spliced.
  • Deletion 47 bp in exon 4 leads to skipping of the 4th exon during splicing and disruption of RNA synthesis and expression of the active enzyme.
  • DNA present in the culture medium is fragmented to a size of 500-1000 bp. hydrodynamic method. This means that in the experiment there are fundamentally no fragments capable of being used for the synthesis of a full-sized product giving a functional enzyme. Elusively small probability that the ligation of the necessary fragments in the correct orientation occurred with the restoration of the open reading frame of the caspase gene and the formation, thereby
  • the claimed method for the treatment of cancer was investigated in the system (ip vivo) on experimental tumors of mice
  • mice of the A / Sp, CBA / Lac, C57BL / 6 and ICR mice were tested for the survival of the mice of the A / Sp, CBA / Lac, C57BL / 6 and ICR mice at the age of 3-4 months of vivarium wiring of the Institute of Cytology and Genetics, SB RAS.
  • Three strains of transplantable tumors were used in the studies: Ehrlich tumor, GA-1 hepatoma, and Lewis tumor.
  • Ehrlich ascites tumor and Lewis lung carcinoma were obtained from the Oncology Research Institute of the TNC SB RAMS.
  • Hepatoma GA-1 was primarily induced in a mouse of the A / He line by ortho-aminoazotoluene. It is maintained in ascites form in A / Sp mice. With subcutaneous or intramuscular inoculation, it grows at the site of inoculation in the form of a solid node and gives multiple metastases to the liver.
  • the source of human DNA was the placenta of healthy mothers, and the DNA of mice was a mixture of organ tissues (liver, kidneys, spleen, thymus). DNA was isolated by the phenolic-free method (laboratory regulations of the ICG SB RAS). This method allows you to get a full genomic DNA with the preservation of those fragments that vivo strongly associated with proteins of the nuclear matrix. The DNA was fragmented with an ultrasonic disintegrator, resulting in a mixture of fragments of 200-6000 bp in size. The preparations were stored in packaged small portions in a freezer at -18 ° C. Before use
  • Table 7 The frequency of structural rearrangements of chromosomes arising in the culture of human diploid cells hTERT-VL, treated for two days with extracellular DNA at a concentration of 300 ⁇ g / ml.
  • SUBSTITUTE SHEET (RULE 26) It was experimentally established that fragmented DNA at a concentration equal to or greater than 300 ⁇ r / ml can lead to inhibition of cell growth and the appearance of chromosomal abnormalities.
  • the concentration of the drug 30 ⁇ g / ml does not have a cytotoxic effect.
  • 5 mg doses are recommended for a single dose. This corresponds to 1 mg of DNA per liter of blood or a working concentration of the drug in the blood of 1 ⁇ g / ml, which is 300 times less than the concentration at which undesirable effects were observed and 30 times lower than exogenous DNA concentrations that did not have cytotoxicity for hTERT-BJ1 human cell culture cells.
  • the desired result is achieved, since it is included in expanding the scope and treatment of patients with cancer, since the method allows you to treat the cause of the disease, namely, to correct the mutations in oncogenes and cancer suppressor genes, as well as to make heterozygous alleles in the cell genes.

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Abstract

L'invention concerne le domaine de la médecine et plus particulièrement des procédés de traitement de patients avec des préparations contenant un matériel génétique. Elle peut s'utiliser pour traiter les malades souffrant de maladies cancéreuses dues à des mutations dans les oncogènes, les gènes d'oncosuppresseurs et à l'homozygatisation totale des allèles de gènes de la cellule qui a subi une mutation cancéreuse. L'invention permet d'élargir le domaine d'application et de traitement des malades souffrant de maladies cancéreuses dues à des mutations dans les oncogènes, les gènes d'oncosuppresseurs ou à l'homozygatisation totale des gènes; selon l'invention, on introduit dans l'organisme du patient des fragments d'un ADN homologique constituant le génome total d'un donneur physiologiquement et génétiquement sain. La quantité d'ADN introduite est égale ou supérieure à celle d'ADN du plasma sanguin et des liquides tissulaires du patient mais sans dépasser la valeur admissible maximale de 30 microgrammes par millilitre.
PCT/RU2006/000144 2006-01-16 2006-03-27 Procede de traitement de maladies cancereuses Ceased WO2007081235A1 (fr)

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EP06769535A EP2002840A4 (fr) 2006-01-16 2006-03-27 Procede de traitement de maladies cancereuses
JP2008551212A JP2009523788A (ja) 2006-01-16 2006-03-27 腫瘍性疾患の治療用製剤

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RU2006100561/14A RU2313349C2 (ru) 2006-01-16 2006-01-16 Способ лечения онкологических заболеваний

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RU2387456C1 (ru) * 2008-08-28 2010-04-27 Михаил Аркадьевич Шурдов Способ лечения онкологических заболеваний
IT1393331B1 (it) * 2009-02-09 2012-04-20 Graal S R L Composizioni orosolubili e/o effervescenti contenenti almeno un sale di s-adenosilmetionina (same)
RU2429019C2 (ru) * 2009-08-03 2011-09-20 Михаил Аркадьевич Шурдов Способ лечения онкологических заболеваний

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EP2002840A4 (fr) 2011-07-13
RU2313349C2 (ru) 2007-12-27
RU2006100561A (ru) 2007-07-27
EP2002840A2 (fr) 2008-12-17

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